Preliminary gastroprotective effect of aqueous <i>Bixa orellana</i> leaf extract in an HCl/ethanol‑induced gastric ulcer rat model
PDF

Keywords

Bixa Orellana
gastric ulcer
HCl/ethanol-induced ulcer
histopathology
gastroprotection
medicinal plant

How to Cite

Mohd Yusof, A. S., Yong, Y. K., Fong, L. Y., Ng, C. T., Mohd Sidik, S., Abdul Moin, S., Omar Fauzee, M. S., Abdullah, M. N. H., & Ahmad, Z. (2026). Preliminary gastroprotective effect of aqueous Bixa orellana leaf extract in an HCl/ethanol‑induced gastric ulcer rat model. Life Sciences, Medicine and Biomedicine, 10(1). https://doi.org/10.28916/lsmb.10.1.2026.278

Abstract

Gastric ulceration develops when aggressive luminal factors overwhelm gastric mucosal defense. Bixa orellana leaves are used traditionally for gastrointestinal complaints, but direct evidence for aqueous B. orellana leaf extract (BOE) in chemically induced gastric ulcer models remains limited. This exploratory study evaluated the effects of aqueous BOE against HCl/ethanol-induced gastric ulceration in rats. Twenty-four male Sprague-Dawley rats were allocated to four groups (n = 6/group): untreated ulcer control, omeprazole 100 mg/kg, BOE 50 mg/kg, and BOE 150 mg/kg. Treatments were administered orally for 7 days. After 24 h fasting, ulcers were induced with 1.0 mL HCl/ethanol, and stomach tissues were collected for macroscopic ulcer-index scoring and hematoxylin and eosin histological assessment. No measured outcome reached statistical significance. The ulcer index showed a non-significant overall difference among groups (Kruskal-Wallis H = 6.57, p = 0.087). Histological scores were also non-significant for congestion (p = 0.458), edema (p = 0.158), inflammatory infiltration (p = 0.126), and necrosis (p = 0.089). Omeprazole and BOE 150 mg/kg showed numerically lower ulcer-index values than the untreated ulcer control, whereas BOE 50 mg/kg remained close to the ulcer control. BOE 150 mg/kg also showed numerically lower edema, inflammatory infiltration, and necrosis scores, but these differences were not statistically confirmed. The present study demonstrates biological trends rather than statistically confirmed gastroprotective efficacy. Aqueous BOE showed a possible dose-related gastroprotective trend, with 150 mg/kg producing the most favorable numerical pattern. Larger dose-response studies with extract standardization, blinded morphometric assessment, ulcer-area and mucus endpoints, and biochemical assays are required before efficacy or mechanistic claims can be made.

PDF

References

Ahmad, S., Israf, D. A., Lajis, N. H., Shaari, K., Mohamed, H., Wahab, A. A., Ariffin, K. T., Hoo, W. Y., Aziz, N. A., Kadir, A. A., Sulaiman, M. R., & Somchit, M. N. (2006). Cardamonin inhibits pro-inflammatory mediators in activated RAW 264.7 cells and whole blood. European Journal of Pharmacology, 538(1-3), 188-194.

https://doi.org/10.1016/j.ejphar.2006.03.070

Ahmad, Z., Ng, C. T., Fong, L. Y., Abu Bakar, N. A., Mohd Hussain, N. H., Ang, K. P., Ee, G. C. L., & Hakim, M. N. (2016). Cryptotanshinone inhibits TNF-alpha-induced early atherogenic events in vitro. The Journal of Physiological Sciences, 66(3), 213-220.

https://doi.org/10.1007/s12576-015-0410-7

Ahmed, O., Nedi, T., Yimer, E. M., Ayalew, W., Shibeshi, W., & Asefa, A. (2022). Evaluation of anti-gastric ulcer activity of aqueous and 80% methanol leaf extracts of Urtica simensis in rats. Metabolism Open, 13, 100155.

https://doi.org/10.1016/j.metop.2021.100155

Alhatim, H., Alshadfan, H., Omar Fauzee, M. S., Zakaria, Z. A., & Hakim, M. N. (2026). Analyzing NSAIDs research trends: A six-year bibliometric study (2018-2023) on directions, themes, and dimensions. Multidisciplinary Reviews, 9(3), 2026130.

https://doi.org/10.31893/multirev.2026130

Almadi, M. A., Lu, Y., Alali, A. A., & Barkun, A. N. (2024). Peptic ulcer disease. The Lancet, 404(10456), 68-81.

https://doi.org/10.1016/S0140-6736(24)00155-7

Alshadfan, H., Jarrar, Q., Omar Fauzee, M. S., Zakaria, Z. A., Yong, Y. K., & Hakim, M. N. (2025). Non-steroidal anti-inflammatory drug-induced gastric ulcers: A review on some current issues. Tropical Journal of Pharmaceutical Research, 24(6), 839-846.

https://doi.org/10.4314/tjpr.v24i6.11

Badr, A. M., El-Orabi, N. F., & Ali, R. A. (2019). The implication of the crosstalk of Nrf2 with NOXs and HMGB1 in ethanol-induced gastric ulcer: Potential protective effect afforded by raspberry ketone. PLOS ONE, 14(8), e0220548.

https://doi.org/10.1371/journal.pone.0220548

Chey, W. D., Howden, C. W., Moss, S. F., Morgan, D. R., Greer, K. B., Grover, S., & Shah, S. C. (2024). ACG clinical guideline: Treatment of Helicobacter pylori infection. The American Journal of Gastroenterology, 119(9), 1730-1753.

https://doi.org/10.14309/ajg.0000000000002968

da Silva, V. R. P., Martins, N. O., dos Santos, C. R., de Oliveira, M. A., Araujo, P. L., Damas, E., & Carneiro, M. (2024). Annatto (Bixa orellana)-based nanostructures for biomedical applications: A systematic review. Pharmaceutics, 16(10), 1275.

https://doi.org/10.3390/pharmaceutics16101275

de Araújo Vilar, D., de Araújo Vilar, M. S., de Lima e Moura, T. F. A., Raffin, F. N., de Oliveira, M. R., Franco, C. F. O., de Athayde-Filho, P. F., Diniz, M. F. F. M., & Barbosa-Filho, J. M. (2014). Traditional uses, chemical constituents, and biological activities of Bixa orellana L.: A review. The Scientific World Journal, 2014, 857292.

https://doi.org/10.1155/2014/857292

Franklin, V. A., Edgar, M., Wadt, N. S. Y., & Bach, E. E. (2023). Aqueous extract from urucum (Bixa orellana L.): Antimicrobial, antioxidant, and healing activity. Porto Biomedical Journal, 8, e183.

https://doi.org/10.1097/j.pbj.0000000000000183

Gamberini, M. T., Skorupa, L. A., Souccar, C., & Lapa, A. J. (1991). Inhibition of gastric secretion by a water extract from Baccharis triptera Mart. Memórias do Instituto Oswaldo Cruz, 86(Suppl. 2), 137-139.

https://doi.org/10.1590/S0074-02761991000600031

Guerrero-Lagunes, L. A., Ruiz-Posadas, L. M., Cadena-Iñiguez, J., Soto-Hernández, R. M., Avendaño-Arrazate, C. H., Aguirre-Medina, J. F., Soto-Mendoza, C., & Aguirre-Cadena, J. F. (2025). The bioprospecting of Bixa orellana L. for the selection of characters with biological activity. Metabolites, 15(2), 115.

https://doi.org/10.3390/metabo15020115

Kamada, T., Satoh, K., Itoh, T., Ito, M., Iwamoto, J., Okimoto, T., Kanno, T., Sugimoto, M., Chiba, T., Nomura, S., & Mizokami, Y. (2021). Evidence-based clinical practice guidelines for peptic ulcer disease 2020. Journal of Gastroenterology, 56(4), 303-322.

https://doi.org/10.1007/s00535-021-01769-0

Mizui, T., & Doteuchi, M. (1983). Effect of polyamines on acidified ethanol-induced gastric lesions in rats. Japanese Journal of Pharmacology, 33(5), 939-945.

https://doi.org/10.1254/jjp.33.939

Molina-Romani, S. P., Bonilla-Rivera, P. E., & de Albuquerque, R. D. D. G. (2023). A review of Bixa orellana L. (annatto) leaves as medicinal resource: Use in the population as complementary medicine, phytotherapeutic action and quality parameters. Natural Resources for Human Health, 3, 277-285.

https://doi.org/10.53365/nrfhh/161027

Ng, C. T., Fong, L. Y., Low, Y. Y., Ban, J., Hakim, M. N., & Ahmad, Z. (2016). Nitric oxide participates in IFN-gamma-induced HUVECs hyperpermeability. Physiological Research, 65(6), 1053-1058.

https://doi.org/10.33549/physiolres.933237

Percie du Sert, N., Hurst, V., Ahluwalia, A., Alam, S., Avey, M. T., Baker, M., Browne, W. J., Clark, A., Cuthill, I. C., Dirnagl, U., Emerson, M., Garner, P., Holgate, S. T., Howells, D. W., Karp, N. A., Lazic, S. E., Lidster, K., MacCallum, C. J., Macleod, M., Pearl, E. J., Petersen, O. H., Rawle, F., Reynolds, P., Rooney, K., Sena, E. S., Silberberg, S. D., Steckler, T., & Würbel, H. (2020). The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLOS Biology, 18(7), e3000410.

https://doi.org/10.1371/journal.pbio.3000410

Raish, M., Ahmad, A., Ansari, M. A., Alkharfy, K. M., Alqahtani, S., Ahmad, S. F., Al-Jenoobi, F. I., Al-Mohizea, A. M., & Ganaie, M. A. (2021). Gastroprotective effect of sinapic acid on ethanol-induced gastric ulcers in rats: Involvement of Nrf2/HO-1 and anti-inflammatory pathways. Frontiers in Pharmacology, 12, 622815.

https://doi.org/10.3389/fphar.2021.622815

Robert, A. (1979). Cytoprotection by prostaglandins. Gastroenterology, 77(4 Pt 1), 761-767.

https://doi.org/10.1016/0016-5085(79)90235-X

Sarkar, K. K., Mitra, T., Hossain, M. M., Aktaruzzaman, M., & Abid, M. A. (2024). Exploring antioxidative, cytotoxic and neuropharmacological insights into Bixa orellana leaves: Experimental and in silico approaches. Heliyon, 10, e27001.

https://doi.org/10.1016/j.heliyon.2024.e27001

Somchit, M. N., Mohamed, N. A., Ahmad, Z., Zakaria, Z. A., Shamsuddin, L., Omar Fauzee, M. S., & Abdul Kadir, A. (2014). Anti-inflammatory and antipyretic properties of Spirulina platensis and Spirulina lonar: A comparative study. Pakistan Journal of Pharmaceutical Sciences, 27(5), 1277-1280.

https://www.pjps.pk/uploads/pdfs/27/5/Paper-22.pdf

Wang, S., Zhang, T., Li, D., & Cao, X. (2025). The global, regional and national burden of peptic ulcer disease attributable to smoking from 1990 to 2021: A population-based study. Preventive Medicine Reports, 51, 103019. https://doi.org/10.1016/j.pmedr.2025.103019

Yong, Y. K., Abdul Kadir, A., Sukardi, S., Abd Hamid, R., Somchit, M. N., & Ahmad, Z. (2011). Bixa orellana leaves extract inhibits bradykinin-induced inflammation through suppression of nitric oxide production. Medical Principles and Practice, 20(2), 142-146.

https://doi.org/10.1159/000319907

Yong, Y. K., Zakaria, Z. A., Abdul Kadir, A., Somchit, M. N., Ee Cheng Lian, G., & Ahmad, Z. (2013a). Chemical constituents and antihistamine activity of Bixa orellana leaf extract. BMC Complementary and Alternative Medicine, 13, 32.

https://doi.org/10.1186/1472-6882-13-32

Yong, Y. K., Sulaiman, N. S., Abdullah, M. N. H., Ee, G. C. L., Zakaria, Z. A., Othman, F., & Ahmad, Z. (2013b). Suppression of serotonin-induced increased vascular permeability and leukocyte infiltration by Bixa orellana leaf extract. BioMed Research International, 2013, 463145.

https://doi.org/10.1155/2013/463145

Yong, Y. K., Chiong, H. S., Somchit, M. N., & Ahmad, Z. (2015). Bixa orellana leaf extract suppresses histamine-induced endothelial hyperpermeability via the PLC-NO-cGMP signaling cascade. BMC Complementary and Alternative Medicine, 15, 356.

https://doi.org/10.1186/s12906-015-0901-3

Yong, Y. K., Somchit, M. N., & Ahmad, Z. (2018). Anti-inflammatory properties of Bixa orellana leaves extract are associated with suppression of bradykinin-induced endothelial hyperpermeability. Pharmacognosy Magazine, 14(Suppl.), S352-S357.

https://phcog.com/article/view/2018/14/57s/s352-s357

Zabidi, Z., Wan Zainulddin, W. N., Mamat, S. S., Shamsahal Din, S., Kamisan, F. H., Yahya, F., Ismail, N. A., Rodzi, R., Hassan, H., Mohtarrudin, N., Somchit, M. N., & Zakaria, Z. A. (2012). Antiulcer activity of methanol extract of Melastoma malabathricum leaves in rats. Medical Principles and Practice, 21(5), 501-503.

https://doi.org/10.1159/000337406

Zhang, S. L., Lollie, T. K., Chen, Z., Narasimhalu, T., & Wang, H. L. (2023). Histopathologic diagnosis of gastritis and gastropathy: A narrative review. Digestive Medicine Research, 6, 9.

https://doi.org/10.21037/dmr-22-42

Zhou, D., Yang, Q., Tian, T., Chang, Y., Li, Y., Duan, L. R., Li, H., & Wang, S. W. (2020). Gastroprotective effect of gallic acid against ethanol-induced gastric ulcer in rats: Involvement of Nrf2/HO-1 signaling and anti-apoptosis role. Biomedicine & Pharmacotherapy, 126, 110075.

https://doi.org/10.1016/j.biopha.2020.110075

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Copyright (c) 2026 Asma Syuhaily Mohd Yusof, Yoke Keong Yong, Lai Yen Fong, Chin Theng Ng, Shiran Mohd Sidik, Saidi Abdul Moin, Mohd Sofian Omar Fauzee, Muhammad Nazrul Hakim Abdullah, Zuraini Ahmad